Development of lensless light-sheet sources and their application for compact selective plane illumination microscopy (SPIM)

We developed a novel method to create a light sheet that require neither laser light nor optical components such as lenses. By coupling a light-emitting diode light to a 10- to 20-μm-thick, 10-mm-long glass sheet, a light sheet having with a thickness of 20 to 40 μm and a width of 20 mm was generated. Our apparatus, HandySPIM, not only generates the light sheet but also holds and moves the sample container. When mounted on the stage of a conventional two-dimensional microscope, HandySPIM transforms it into a three-dimensional (3D) microscope, achieving 3D imaging of biological and pathological specimens with a resolution of 5 μm. We also discuss LUCID, a tissue-clearing reagent that enhances transparency, allows for various staining techniques, and preserves specimens for up to 10 years. The combination of LUCID and HandySPIM represents a significant advancement in 3D imaging, offering transformative potential in the fields of biology and pathology.